Methods for qPCR gene expression profiling applied to 1440 lymphoblastoid single cells

Kenneth J Livak, Quin F Wills, Alex J Tipping, Krishnalekha Datta, Rowena Mittal, Andrew J Goldson, Darren W Sexton, Chris C Holmes

Research output: Contribution to journalArticle

99 Citations (Scopus)

Abstract

The stochastic nature of generating eukaryotic transcripts challenges conventional methods for obtaining and analyzing single-cell gene expression data. In order to address the inherent noise, detailed methods are described on how to collect data on multiple genes in a large number of single cells using microfluidic arrays. As part of a study exploring the effect of genotype on Wnt pathway activation, data were collected for 96 qPCR assays on 1440 lymphoblastoid cells. The description of methods includes preliminary data processing steps. The methods used in the collection and analysis of single-cell qPCR data are contrasted with those used in conventional qPCR.
Original languageEnglish
Pages (from-to)71-9
Number of pages9
JournalMethods
Volume59
Issue number1
DOIs
Publication statusPublished - Jan 2013

Keywords

  • Cell Line
  • Data Interpretation, Statistical
  • Gene Expression Profiling
  • Humans
  • Limit of Detection
  • Lymphoid Progenitor Cells
  • Real-Time Polymerase Chain Reaction
  • Reverse Transcriptase Polymerase Chain Reaction
  • Single-Cell Analysis
  • Wnt Signaling Pathway

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